金藤清痹颗粒通过调节免疫微环境对类风湿关节炎大鼠的干预作用  

Effects of Jinteng Qingbi Granules on rheumatoid arthritis rats through intervention of immune microenvironment

作　　者：唐今扬[1] 周彩云[1] 王鑫[1] 马芳 潘峥[1] 韩淑花[1] 李斌[1] 杜丽妍[1] 程国良 房定亚[1] TANG Jin-yang;ZHOU Cai-yun;WANG Xin;MA Fang;PAN Zheng;HAN Shu-hua;LI Bin;DU Li-yan;CHENG Guo-liang;FANG Ding-ya(Department of Rheumatology,Xiyuan Hospital of China Academy of Chinese Medical Sciences,Beijing 100091,China;State Key Laboratory of Generic Manufacture Technology for Chinese Traditional Medicine,Linyi 276000,China)

机构地区：[1]中国中医科学院西苑医院风湿病科,北京100091 [2]中药制药共性技术国家重点实验室,山东临沂276000

出　　处：《中成药》2022年第11期3459-3468,共10页Chinese Traditional Patent Medicine

基　　金：中国中医科学院科技创新工程项目(CI2021A01512);国家重点研发计划项目(2018YFC1705502)。

摘　　要：目的研究金藤清痹颗粒通过调节免疫微环境对类风湿关节炎大鼠的干预作用及其机制。方法通过注射牛Ⅱ型胶原和弗氏不完全佐剂建立胶原诱导型关节炎(CIA)大鼠模型,造模成功后随机分为模型组、甲氨蝶呤组、雷公藤多苷组及金藤清痹颗粒低、中、高剂量组,每组10只,另以10只正常大鼠作为正常组,给药组灌胃给予相应剂量药物,模型组和正常组灌胃给予等体积0.9%氯化钠注射液。每7 d进行1次足底厚度检测和关节指数评分,HE染色观察膝关节滑膜病理改变,免疫组化检测滑膜CD68、CD86、CD206表达,流式细胞术检测外周血中辅助性T细胞17(Th17)和调节性T细胞(Treg)比例,Western blot法检测TLR4/NF-κB信号通路蛋白表达,RT-qPCR法检测TLR4、IKKβmRNA表达。结果金藤清痹颗粒与甲氨蝶呤、雷公藤多苷均能减轻CIA大鼠四肢关节肿胀(P<0.05),降低M1/M2型巨噬细胞比例、Th17/Treg细胞比例、TLR 4 mRNA及TLR4、p-IκBα、细胞核p65蛋白表达(P<0.05,P<0.01),升高IKKβ、胞质p65蛋白及IKKβmRNA表达(P<0.05,P<0.01),并呈剂量依赖性。结论金藤清痹颗粒能够通过影响M1/M2型巨噬细胞极化,调节Th17/Treg细胞平衡,调控TLR4/NF-κB信号通路表达,改善免疫微环境,从而缓解类风湿关节炎大鼠关节肿胀症状。AIM To investigate the effects and mechanism of Jinteng Qingbi Granules on rheumatoid arthritis rats through intervention of the immune microenvironment.METHODS After the successful establishment of rat models of collagen-induced arthritis(CIA)by injection of both bovine collagenⅡand incomplete Freund’s adjuvant,the rats were randomly divided into the model group,methotrexate group,tripterygium polyglycoside group,and low dose,medium dose and high dose Jinteng Qingbi Granules groups,with 10 rats in each group and another 10 normal rats in the normal group.Gavage administration of corresponding dose of drug was given to the drug groups,while gavage feeding of the same volume of 0.9%sodium chloride injection was dosed upon the model group and normal group.Every 7 days,the rats had their plantar thickness measured and joint index scored;their pathological changes of synovium of knee joint observed by HE staining;their expressions of CD68,CD86 and CD206 in synovium detected by immunohistochemistry;their proportions of helper T cells 17(Th17)and regulatory T cells(Treg)in peripheral blood detected by flow cytometry;their TLR4/NF-κB signaling pathway proteins expressions detected by Western blot;and their mRNA expressions of TLR 4 and IKKβdetected by RT-qPCR.RESULTS Jinteng Qingbi Granules,methotrexate and tripterygium polyglycosides dose-dependently reduced the swelling of the limbs and joints of CIA rats(P<0.05);reduced the proportion of M1/M2 macrophages,the proportion of Th17/Treg cells,the expressions of TLR 4 mRNA and TLR4,p-IκBαand nuclear p65 proteins(P<0.05,P<0.01);and increased the expressions of IKKβand cytoplasmic p65 proteins as well as IKKβmRNA(P<0.05,P<0.01).CONCLUSION Jinteng Qingbi Granules can improve the immune microenvironment to control rheumatoid arthritis joint swelling in rats by affecting the polarization of M1/M2 macrophages,regulating Th17/Treg cells balance and the expression of TLR4/NF-κB signaling pathway.

关 键 词：金藤清痹颗粒 类风湿关节炎 巨噬细胞 M1/M2极化 Th17/Treg平衡 TLR4/NF-κB信号通路 免疫微环境 

分 类 号：R285.5[医药卫生—中药学]